High-content screens reveal residents and effectors of the nucleus–mitochondria contact. (A) Illustration of the high-content screen directed at finding resident proteins of the nucleus–mitochondria contact site. The reporter (NSG1-VN/TOM70-VC) was integrated into a library of ∼6,000 yeast strains, each harboring an overexpressed and mCherry-tagged version of a different yeast protein. Strains were imaged using automated microscopy, and images were manually examined to identify proteins that colocalize, either fully or partially, with the reporter. (B) List of all proteins that either fully (left) or partially (right) colocalized with the reporter, organized by alphabetical order. The proteins shown in the representative images are marked in bold. Scale bars, 5 µm. For a complete list of all proteins and their descriptions, see Table S1. (C) Illustration of a screen aimed at identifying effectors of the nucleus–mitochondria contact site. The reporter (NSG1-VC/TOM70-VN) was integrated into all 57 hits from the primary screen (shown in B). The effect of their overexpression on the reporter was inspected and 12 hits were found. A representative image shows the protein marked in bold out of the full list of hits. Scale bar, 5 µm.